The long-term objective of this research is to facilitate studies aimed at improving our understanding of the role of the epididymis in sperm maturation and how the process might be regulated by contraceptive agents. Additional experimental techniques which permit direct study of the epididymal epithelium and regional differences in its physiology are essential to enable fundamental studies prerequisite to applied studies aimed at manipulating epididymal physiology. Therefore, this research proposal is keyed on developing and perfecting techniques to (a) separate living epididymal epithelial cell types and defining optimum conditions for their culture and (b) culturing tubule fragments from specific epididymal regions. A velocity sedimentation technique, possibly augmented by density techniques, will be employed to separate cells from the caput epididymidis. Fragments of ductus epididymidis will be isolated by enzymatic digestion or acid maceration from the caput or cauda epididymidis. To determine optimum culture conditions for epididymal cells and tubule fragments, the effects of incubation temperature, atmosphere, pH, culture nutrients, vitamins and hormones will be evaluated. Culture techniques will be evaluated on the basis of ultrastructure and histochemical comparisons of cells sampled before and after culture. When suitable culture techniques have been established, biochemical and metabolic studies will be initiated to evaluate functional differences between cell types or epididymal regions.